重组Anti-VCP抗体[EPR3307(2)] (ab109240)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3307(2)] to VCP
- Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-VCP抗体[EPR3307(2)]
参阅全部 VCP 一抗 -
描述
兔单克隆抗体[EPR3307(2)] to VCP -
宿主
Rabbit -
经测试应用
适用于: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- MCF-7 cell lysate, HeLa whole cell lysate (ab150035), A549 cell lysate; SH-SY5Y cell lysate; human breast carcinoma tissue; HeLa cells
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR3307(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab109240于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (1) |
1/10000. Predicted molecular weight: 89 kDa.
For unpurifed use at 1/10000-1/50000. |
IP |
1/10 - 1/100.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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ICC/IF |
1/500.
For unpurified use at 1/100 - 1/250. |
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Flow Cyt (Intra) |
1/300.
For unpurified use at 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
1/10000. Predicted molecular weight: 89 kDa. For unpurifed use at 1/10000-1/50000. |
IP
1/10 - 1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
ICC/IF
1/500. For unpurified use at 1/100 - 1/250. |
Flow Cyt (Intra)
1/300. For unpurified use at 1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
Necessary for the fragmentation of Golgi stacks during mitosis and for their reassembly after mitosis. Involved in the formation of the transitional endoplasmic reticulum (tER). The transfer of membranes from the endoplasmic reticulum to the Golgi apparatus occurs via 50-70 nm transition vesicles which derive from part-rough, part-smooth transitional elements of the endoplasmic reticulum (tER). Vesicle budding from the tER is an ATP-dependent process. The ternary complex containing UFD1L, VCP and NPLOC4 binds ubiquitinated proteins and is necessary for the export of misfolded proteins from the ER to the cytoplasm, where they are degraded by the proteasome. The NPLOC4-UFD1L-VCP complex regulates spindle disassembly at the end of mitosis and is necessary for the formation of a closed nuclear envelope (By similarity). Regulates E3 ubiquitin-protein ligase activity of RNF19A. -
疾病相关
Defects in VCP are the cause of inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD) [MIM:167320]; also known as muscular dystrophy, limb-girdle, with Paget disease of bone or pagetoid amyotrophic lateral sclerosis or pagetoid neuroskeletal syndrome or lower motor neuron degeneration with Paget-like bone disease. IBMPFD features adult-onset proximal and distal muscle weakness (clinically resembling limb girdle muscular dystrophy), early-onset Paget disease of bone in most cases and premature frontotemporal dementia. -
序列相似性
Belongs to the AAA ATPase family. -
翻译后修饰
Phosphorylated by tyrosine kinases in response to T-cell antigen receptor activation (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
ISGylated. -
细胞定位
Cytoplasm > cytosol. Nucleus. Present in the neuronal hyaline inclusion bodies specifically found in motor neurons from amyotrophic lateral sclerosis patients. Present in the Lewy bodies specifically found in neurons from Parkinson disease patients. - Information by UniProt
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数据库链接
- Entrez Gene: 7415 Human
- Entrez Gene: 269523 Mouse
- Entrez Gene: 116643 Rat
- Omim: 601023 Human
- SwissProt: P55072 Human
- SwissProt: Q01853 Mouse
- SwissProt: P46462 Rat
- Unigene: 529782 Human
see all -
别名
- 15S Mg(2+) ATPase p97 subunit antibody
- 15S Mg(2+)-ATPase p97 subunit antibody
- ALS14 antibody
see all
图片
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All lanes : Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates)
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
Lane 5 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 6 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 89 kDa
Observed band size: 89 kDaBlocking and diluting buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling VCP with purified ab109240 at 1:250 dilution (1.4 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunofluorescent staining of HeLa cells using unpurified ab109240 at 1/100 dilution.
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ab109240 (purified) at 1:20 dilution (2ug) immunoprecipitating VCP in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab109240 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109240 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Intracellular Flow Cytometry analysis of HL-60 (Human acute promyelocytic leukemia promyeloblast) cells labeling VCP with purified ab109240 at 1/300 dilution (1 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488) secondary antibody was used at 1/2000 dilution. Isotype control - 90% methanol. Unlabeled control - Rabbit monoclonal IgG (Black).Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling VCP with Purified ab109240 at 1:500 dilution (0.7μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab109240 at 1/100 dilution.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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All lanes : Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution (unpurified)
Lane 1 : MCF7 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : A549 cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg/ml per lane.
Predicted band size: 89 kDa -
Overlay histogram showing HL60 cells stained with unpurified ab109240 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109240, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HL60 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (24)
ab109240 被引用在 24 文献中.
- Liu F et al. Proximity proteomics of C9orf72 dipeptide repeat proteins identifies molecular chaperones as modifiers of poly-GA aggregation. Acta Neuropathol Commun 10:22 (2022). PubMed: 35164882
- Wang M et al. Pharmacological activation of ATF6 remodels the proteostasis network to rescue pathogenic GABAA receptors. Cell Biosci 12:48 (2022). PubMed: 35477478
- Prophet SM et al. p97/UBXD1 Generate Ubiquitylated Proteins That Are Sequestered into Nuclear Envelope Herniations in Torsin-Deficient Cells. Int J Mol Sci 23:N/A (2022). PubMed: 35563018
- Wrobel L et al. Compounds activating VCP D1 ATPase enhance both autophagic and proteasomal neurotoxic protein clearance. Nat Commun 13:4146 (2022). PubMed: 35842429
- Yang H et al. GINS1 promotes the proliferation and migration of glioma cells through USP15-mediated deubiquitination of TOP2A. iScience 25:104952 (2022). PubMed: 36065190